To investigate the effects of Snail1 gene silence on the expression of tight junction proteins and the migration ability of Hep-2 cells, Hep-2 cells were transfected with plasmids which is containing the shRNA of Snail1 gene, and cultured till the cells could be passaged stably (named Sh-snail1 cells). The expression of tight junction proteins (ZO-1, Occludin, Claudin-5) were detected by Western blot. The migration ability of Sh-snail1 cells was investigated by wound healing assay, and the protein expression of members of RhoGTPase family (RhoA, Cdc42) was detected by Western blot, which is closely related to the migration ability. Our results showed that the expression of tight junction proteins (ZO-1, Occludin, Claudin-5) was significantly increased; the migration ability of Sh-snail1 cell was inhibited; the expression of RhoA and Cdc42 was downregulated. All of these indicated that silencing the gene of Snail1 in Hep-2 cells can up-regulate the expression of tight junction proteins and down regulate the expression of Cdc42 and RhoA, and further inhibit the migration of Hep-2 cells. Furthermore, opening of the tight junctions between cells and the stronger migration ability of cancer cells are important processes in cancer metastasis. It is confirmed that the Snail1 gene is closely related to the two processes, providing an experimental basis for targeted therapy of laryngeal squamous cell carcinoma.
本文为研究 Snail1 基因对喉鳞癌细胞株 Hep-2 紧密连接蛋白表达及迁移能力的影响，将含有 Snail1 基因的短发夹 RNA（Sh-RNA）的质粒转染喉鳞癌细胞株 Hep-2，培养出可稳定传代的 Snail1 基因沉默的细胞（命名为 Sh-snail1 细胞）。课题组以蛋白免疫印迹技术检测紧密连接蛋白 ZO-1、Occludin、Claudin-5 的表达是否发生变化。然后设计伤口愈合实验检测其迁移能力，再以蛋白免疫印迹技术检测与细胞迁移能力密切相关的 RhoGTP 酶家族重要成员 RhoA、Cdc42 蛋白的表达变化。结果表明，Sh-snail1 细胞紧密连接关键蛋白 ZO-1、Occludin、Claudin-5 表达明显上调，迁移能力明显减弱，且 RhoA、Cdc42 蛋白表达下调。该研究表明，喉鳞癌 Hep-2 细胞通过下调 Snail1 基因的表达使细胞间连接更加紧密，同时抑制 Hep-2 细胞的迁移能力，下调 RhoA、Cdc42 蛋白的表达。本文研究结果证实， Snail1 基因与 Hep-2 细胞转移过程中的细胞间紧密连接打开以及细胞迁移能力增强密切相关，为靶向治疗喉鳞癌提供分子机制的研究依据。.
Keywords: Hep-2 cell; Snail1; cell migration; gene silence; tight junction.